LOCUS       sequence-310850-        5713 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of mcherry:MFF in cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5713)
  AUTHORS   Bertolin G, Bulteau AL, Alves-Guerra MC, Burel A, Lavault MT, Gavard
            O, Le Bras S, Gagne JP, Poirier GG, Le Borgne R, Prigent C, Tramier 
            M
  TITLE     Aurora kinase A localises to mitochondria to control organelle 
            dynamics and energy production.
  JOURNAL   eLife. 2018 Aug 2;7. pii: 38111. doi: 10.7554/eLife.38111.
  PUBMED    30070631
REFERENCE   2  (bases 1 to 5713)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "eLife."; 
            date: "2018-08-2"; volume: "7. pii: 38111. doi"; pages: " 
            10.7554/eLife.38111"
COMMENT     Sequence Label: pmCherry C1 MFF
FEATURES             Location/Qualifiers
     source          1..5713
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     276..295
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(594..613)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    638..685
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      1014..1602
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1503..1522
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        1777..2080
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2081..2284
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2235..2255
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      3379..3388
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3385..4092
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(3530..3548)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     3798..3817
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    4217..4338
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4254..4273)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4308..4327
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4345..4800)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4482..4503)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4694..4713
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4827..4931
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4933..5290
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4933..5124
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4956..4976)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5141..5276
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             join(5325..5713,1..406)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             join(5325..5713,1..406)
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(5379..5398)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
ORIGIN
        1 ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc
       61 cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact
      121 gttcgccagg ctcaaggcga gcatgcccga cggcgaggat ctcgtcgtga cccatggcga
      181 tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg
      241 ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga
      301 agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga
      361 ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg
      421 ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga ttccaccgcc
      481 gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc
      541 cagcgcgggg atctcatgct ggagttcttc gcccacccta gggggaggct aactgaaaca
      601 cggaaggaga caataccgga aggaacccgc gctatgacgg caataaaaag acagaataaa
      661 acgcacggtg ttgggtcgtt tgttcataaa cgcggggttc ggtcccaggg ctggcactct
      721 gtcgataccc caccgagacc ccattggggc caatacgccc gcgtttcttc cttttcccca
      781 ccccaccccc caagttcggg tgaaggccca gggctcgcag ccaacgtcgg ggcggcaggc
      841 cctgccatag cctcaggtta ctcatatata ctttagattg atttaaaact tcatttttaa
      901 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
      961 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     1021 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     1081 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     1141 gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac
     1201 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     1261 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     1321 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     1381 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     1441 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     1501 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     1561 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     1621 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc
     1681 cctgattctg tggataaccg tattaccgcc atgcattagt tattaatagt aatcaattac
     1741 ggggtcatta gttcatagcc catatatgga gttccgcgtt acataactta cggtaaatgg
     1801 cccgcctggc tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc
     1861 catagtaacg ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac
     1921 tgcccacttg gcagtacatc aagtgtatca tatgccaagt acgcccccta ttgacgtcaa
     1981 tgacggtaaa tggcccgcct ggcattatgc ccagtacatg accttatggg actttcctac
     2041 ttggcagtac atctacgtat tagtcatcgc tattaccatg gtgatgcggt tttggcagta
     2101 catcaatggg cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga
     2161 cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa
     2221 ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct atataagcag
     2281 agctggttta gtgaaccgtc agatccgcta gcgctaccgg wckcmrmymt sgwgwgmwrr
     2341 gkmraggaac aagcagtgac acatcactag gaagggtgag cagggcagca tttccttctc
     2401 ccactgctgc tgagatggca gaaattagtc gaattcagta cgaaatggaa tatactgaag
     2461 gcattagtca gcgaatgagg gtcccagaaa agttaaaagt agcaccgcca aacgctgacc
     2521 tggaacaagg attccaagaa ggagttccaa atgctagtgt gataatgcaa gttccggaga
     2581 ggattgttgt agcaggaaat aatgaagatg tttcattttc aagaccagca gatcttgacc
     2641 ttattcagtc aactcccttt aaacccctgg cactgaaaac accacctcgt gtacttacgc
     2701 tgagtgaaag accactagat tttctggatt tagaaagacc tcctacaacc cctcaaaatg
     2761 aagaaatccg agcagttggc agactaaaaa gagagcggtc tatgagtgaa aatgctgttc
     2821 gccaaaatgg acagctggtc agaaatgatt ctctgtggca cagatcagat tctgccccaa
     2881 gaaataaaat ttcaaggttc caggcaccga tttctgcacc ggagtacact gtgacaccat
     2941 cgccacaaca ggctcgggtc tgtcctcccc atatgttacc tgaagatgga gctaatcttt
     3001 cctctgctcg tggcattttg tcgcttatcc agtcttctac tcgtagggca taccagcaga
     3061 tcttggatgt gctggatgaa aatcgcagac ctgtgttgcg tggtgggtct gctgccgcca
     3121 cttctaatcc tcatcatgac aacgtcaggt atggcatttc aaatatagat acaaccattg
     3181 aaggaacgtc agatgacctg actgttgtag atgcagcttc actaagacga cagataatca
     3241 aactaaatag acgtctacaa cttctggaag aggagaacaa agaacgtgct aaaagagaaa
     3301 tggtcatgta ttcaattact gtagctttct ggctgcttrr twkmkkgmwc ygkywkmkcc
     3361 gctagsgmtn naccggtcgc caccatggtg agcaagggcg aggaggataa catggccatc
     3421 atcaaggagt tcatgcgctt caaggtgcac atggagggct ccgtgaacgg ccacgagttc
     3481 gagatcgagg gcgagggcga gggccgcccc tacgagggca cccagaccgc caagctgaag
     3541 gtgaccaagg gtggccccct gcccttcgcc tgggacatcc tgtcccctca gttcatgtac
     3601 ggctccaagg cctacgtgaa gcaccccgcc gacatccccg actacttgaa gctgtccttc
     3661 cccgagggct tcaagtggga gcgcgtgatg aacttcgagg acggcggcgt ggtgaccgtg
     3721 acccaggact cctccctgca ggacggcgag ttcatctaca aggtgaagct gcgcggcacc
     3781 aacttcccct ccgacggccc cgtaatgcag aagaagacca tgggctggga ggcctcctcc
     3841 gagcggatgt accccgagga cggcgccctg aagggcgaga tcaagcagag gctgaagctg
     3901 aaggacggcg gccactacga cgctgaggtc aagaccacct acaaggccaa gaagcccgtg
     3961 cagctgcccg gcgcctacaa cgtcaacatc aagttggaca tcacctccca caacgaggac
     4021 tacaccatcg tggaacagta cgaacgcgcc gagggccgcc actccaccgg cggcatggac
     4081 gagctgtaca agtccggact cagatctcga gtgatgagya rmsrwacman caktkryasa
     4141 kswyttactt gctttaaaaa acctcccaca cctccccctg aacctgaaac ataaaatgaa
     4201 tgcaattgtt gttgttaact tgtttattgc agcttataat ggttacaaat aaagcaatag
     4261 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa
     4321 actcatcaat gtatcttaac gcgtaaattg taagcgttaa tattttgtta aaattcgcgt
     4381 taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc aaaatccctt
     4441 ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg aacaagagtc
     4501 cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat cagggcgatg
     4561 gcccactacg tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac
     4621 taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag ccggcgaacg
     4681 tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag
     4741 cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt
     4801 caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
     4861 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
     4921 aaaggaagag tcctgaggcg gaaagaacca gctgtggaat gtgtgtcagt tagggtgtgg
     4981 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
     5041 aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     5101 caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc
     5161 cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg cagaggccga
     5221 ggccgcctcg gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg
     5281 cttttgcaaa gatcgatcaa gagacaggat gaggatcgtt tcgcatgatt gaacaagatg
     5341 gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac
     5401 aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg
     5461 ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaagac gaggcagcgc
     5521 ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg
     5581 aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc
     5641 accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc
     5701 ttgatccggc tac
//
